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Biomedical subjects

R S Jeyendran

Publications and source records attributed to R S Jeyendran.

At least 37 records · Page 2Linked to original sources

Hydrogen hexachloroplatinate induces the acrosome reaction in human spermatozoa.

Human spermatozoa from healthy donors (n = 7) were washed in Tyrode's medium containing 0.6 mg/ml freeze-dried egg yolk, filtered through glass wool and exposed to 0.5, 5, 50, 500 or 1000 microM hydrogen hexachloroplatinate. Viability, membrane integrity and the acrosome reaction were examined using trypan blue exclusion, hypo-osmotic swelling test and fluoresceinated Pisum sativum agglutinin, respectively, after incubation for 3 or 6 h at 37 degrees C. While sperm motility, viability and membrane integrity were not affected by the platinum compound after incubation for 3 h, the number of acrosome-reacted spermatozoa increased from 16.0 +/- 6.4% (control, mean +/- SEM) to 21.0 +/- 3.3 (0.5 microM), 22.3 +/- 4.3% (5 microM), 28.0 +/- 4.3% (50 microM, p < 0.01), 29.3 +/- 3.9% (500 microM, p < 0.01) and 43.9 +/- 7.4% (1 mM, p < 0.001); a further increase was detected after incubation for 6 h. However, the percentages of dead and immotile spermatozoa and those with defective membranes were also higher, suggesting that the acrosome reaction was caused by degenerative processes after long-term incubation. In conclusion, hydrogen hexachloroplatinate does not affect sperm motility, membrane integrity or viability, but it does induce the acrosome reaction after incubation for 3 h before cytotoxic effects are measurable. Similar effects of halide salts of platinum on receptor-mediated exocytosis have been described in other cells such as mast cells and basophils in vivo and in vitro.

Acrosome↗

Variation in antisperm antibody results using different assays.

PROBLEM: The detection of various types of antisperm antibodies (ASA) in the serum varies among different assays. This variation may influence the diagnosis and management of infertile couples who are tested for such immunologic factors. This prospective study was conducted to determine the variation in the results of ASA as measured by the sperm immobilization (SI), sperm agglutination (SA), and the indirect immunobead (IB) assays. METHOD: The sera of 79 patients that tested positive for ASA by at least one of the assays listed above were concurrently tested with all three assays. RESULTS: Using an individual ASA assay, 66 (84%), 26 (33%) or 36 (46%) of sera tested positive by the SA, SI, or IB assays, respectively. However, using a combination of assays, 67 (85%), 78 (99%) or 40 (51%) of sera tested positive using either the SA+SI, SA+IB or SI+IB assays, respectively. CONCLUSION: These findings suggest that the utilization of different assays to detect ASA may detect sera that are positive for ASA with more reliability than single assay testing.

Agglutination Tests↗

Effects of urine on the functional quality of human spermatozoa.

OBJECTIVE: To determine the effect of urine on sperm function and to elucidate the reason by which it may be deleterious. DESIGN: Prospective controlled in vitro study. INTERACTIONS: Sperm were exposed to various concentrations of pooled urine in Ham's F-10 Medium supplemented with 3.5 g% bovine serum albumin (GIBCO, Grand Island, NY) and sperm exposed to Ham's F-10 Medium with pH and osmolality adjusted to mimic that of the urine (pH 5.4; 520 mOsm). MAIN OUTCOME MEASURE: Sperm motility and the ability of sperm to penetrate and migrate into mucus and to penetrate hamster oocytes were determined. RESULTS: All sperm variables studied were significantly decreased when the sperm were exposed to > or = 40% urine. Potential of hydrogen- and osmolality-adjusted Ham's F-10 did not significantly affect the sperm function. CONCLUSION: Urine is deleterious to sperm function and may involve other biochemical factors that are unrelated to pH and osmolality of urine.

Animals↗

Egg yolk enhances the receptiveness of zona pellucida to sperm binding.

PURPOSE: Our purpose was to test whether treatment of zona pellucida with egg yolk enhances its receptiveness to sperm binding. Hams F-10 supplemented with 0.06 g% freeze-dried egg yolk (TM) was evaluated using the hemizona assay. Ham's F-10 supplemented with 7.5% human serum served as the control medium (CM). Salt-stored zona pellucida were bisected and each matched hemizona processed in either TM or CM for the preincubation study or both hemizona processed in CM for the coincubation study. TEST yolk-treated sperm from 10 ejaculates were then incubated with the hemizona preincubated in TM and CM but coincubated in CM or coincubated with hemizona in TM and CM. RESULTS: A significantly (P < 0.02) higher number of tightly bound sperm and hemizona index for the hemizona either preincubated or coincubated in TM compared to the respective hemizonae in CM were observed.

Egg Yolk↗

Effect of preincubation of human spermatozoa in milk on penetration of zona-free hamster oocytes: comparison to TEST yolk.

PURPOSE: To investigate the effect of preincubation of human sperm in milk on their ability to penetrate zona-free hamster oocytes, 27 ejaculates were studied. Each ejaculate was divided into two portions and incubated in either milk or TEST yolk (TESTY) for 22-24 hr at 5 degrees C prior to processing for the sperm penetration assay. RESULTS: Spermatozoa preincubated in milk penetrated a significantly (P < 0.05) higher percentage of oocytes (mean +/- SE; 74.4 +/- 6.3) than spermatozoa preincubated in TESTY (66.8 +/- 6.6). Statistically, no significant differences were noted for penetration index or postincubation sperm motility between the two media. CONCLUSION: Preincubation of sperm in milk therefore appears to enhance their ability to penetrate zona-free hamster oocytes, compared to TESTY preincubation. Since milk is natural, simple, inexpensive, readily available, and can be easily processed, it should be the medium of choice for sperm preincubation.

Animals↗

The effect of preincubation of human spermatozoa in milk on sperm penetration into zona-free hamster oocytes and on sperm binding to the human zona pellucida.

OBJECTIVE: To determine whether preincubation of sperm in milk enhances the outcome of sperm penetration assay (SPA) and hemizona assay (HZA) when compared with preincubation in TES and Tris (TEST)-yolk. DESIGN: Sperm penetration assay and HZA were performed on milk and TEST-yolk preincubated spermatozoa. INTERVENTIONS: Ejaculates were washed and divided into two equal aliquots. An equal volume of heat-inactivated cow's milk (95 degrees C, 10 minutes) was added to one aliquot and an equal volume of TEST-yolk was added to the other aliquot. Both sperm mixtures were cooled slowly to 5 degrees C and incubated for 2 hours. They were then washed twice (400 x g for 8 minutes) with culture medium at 37 degrees C and processed for SPA and HZA. MAIN OUTCOME MEASURES: Percentage of oocytes penetrated and the penetration index for the SPA. The number of sperm bound to hemizona and hemizona index for the HZA. RESULTS: Sperm preincubated in milk yielded significantly higher SPA results and hemizona index when compared with TEST-yolk. The penetration index and the number of sperm bound were high but were not statistically significant. CONCLUSION: Preincubation of sperm in milk enhances the SPA and HZA outcome when compared with TEST-yolk.

Animals↗

Comparison of the methods of artificial insemination on the incidence of conception in single unmarried women.

OBJECTIVE: To compare pregnancy rates after intrauterine insemination (IUI) versus pericervical insemination in absolute male factor infertility using each patient as her own control. DESIGN: Ovulatory women with patent fallopian tubes without male partners were alternately inseminated with cryopreserved donor semen using either IUI or pericervical insemination techniques. A total of 81 cycles, which included up to 4 cycles per patient were performed. In this manner a comparison between the efficacy of each method could be evaluated. SETTING: The donor insemination program at the Center For Assisted Reproduction at Northwestern University Medical School. PATIENTS: Twenty-six single, healthy, unmarried women with patent fallopian tubes and < 40 years of age without male partners (absolute male factor infertility). MAIN OUTCOME MEASURES: Positive quantitative serum subunit of human chorionic gonadotropin followed by the presence of an intrauterine gestational sac seen by transvaginal ultrasonography. RESULTS: Fourteen (54%) of 26 patients conceived including two (14%) miscarriages within four insemination cycles. Seven (17.5%) patients after IUI, and 7 (17.1%) patients after pericervical insemination conceived. The pregnancy rates were similar regardless of the order of insemination method. CONCLUSION: These findings reveal that there is no statistical difference in the pregnancy outcome between these two methods of insemination in absolute male factor infertility.

Adult↗

The straw swim-up: a simple method to evaluate the effects of additives, media, or toxicants on sperm motility.

We describe a new method for evaluating potential toxicants, spermicides, and motility enhancers. Cryopreservation straws filled with media plus additive are emersed below the surface of an unprocessed donor ejaculate. After incubation, the sperm found within the first 4 cm of the straw are expelled and analyzed. The effects of media containing glycerol and albumin were examined. Survivability and sperm migration into the media occur independently and can be differentiated in this assay. This method allows for the use of unprocessed semen and includes an evaluation of the semen and media interaction.

Cell Survival↗

Quality assurance for sperm concentration using latex beads.

OBJECTIVE: To provide a simple, universally applicable method of quality assurance for sperm counting, thereby reducing intercounting chamber variation. DESIGN: By using a known concentration of latex beads, the sperm:bead ratio can be used to calculate the actual sperm count. MAIN OUTCOME MEASURES: The mean sperm and bead counts were determined in both a Spot-lite hemocytometer (Baxter Diagnostics, McGaw Park, IL) and a Makler chamber (Polymedco Inc., Yorktown, NY) from 21 different ejaculates mixed with a known concentration of beads. The hemocytometer chamber was used as the standard counting chamber because it consistently yielded a low variation in sperm count. The adjusted sperm concentration of the Makler chamber was calculated using the following formula [hemocytometer beads]/[Makler beads] x [Makler sperm]. RESULTS: Observed mean +/- SD sperm counts were significantly different between the hemocytometer chamber (110.6 +/- 66.2 x 10(6)/mL) and Makler chamber (173.3 +/- 103.5 x 10(6)/mL). However, calculated Makler chamber sperm counts (118.1 +/- 76.1 x 10(6)/mL) was not statistically different from observed hemocytometer sperm counts. CONCLUSION: This novel approach to sperm counting using a known concentration of latex beads as a reference material can be used to reduce variation in sperm counting between observers, counting chambers, and possibly computerized sperm analyzers.

Equipment and Supplies↗

Effects of peritoneal fluids from patients with endometriosis on capacitated spermatozoa.

OBJECTIVE: To determine the effects of peritoneal fluid (PF) from patients with endometriosis on capacitated sperm. DESIGN: Capacitated donor sperm were allowed to migrate into straws filled with pooled PF from patients with unexplained infertility without endometriosis (n = 4) and those with mild (n = 4) and moderate-severe (n = 4) endometriosis, respectively, for 3 hours. MAIN OUTCOME MEASURES: Sperm motility, sperm membrane integrity, and acrosomal loss were determined. RESULTS: Statistically, no significant differences were found among the various sperm parameters determined. CONCLUSION: Peritoneal fluid from patients with endometriosis does not appear to adversely affect capacitated spermatozoa.

Acrosome↗

Synergistic effect of TEST--yolk buffer treatment and glass wool filtration of spermatozoa on the outcome of the hamster oocyte penetration assay.

Enhanced penetration of zona-free hamster oocytes occurs after spermatozoa have either been treated with TEST--yolk buffer or processed by glass wool filtration. The present study was conducted to determine if a synergistic effect on fertilizing ability could be achieved by combining these two treatments. Ejaculates (n = 16) were treated with TEST--yolk buffer (TYB) and divided into two aliquots. One aliquot was kept as control and the other aliquot was filtered through glass wool prior to the sperm penetration assay. The TYB--glass wool filtered spermatozoa penetrated a significantly higher percentage of oocytes than the TYB treated spermatozoa.

Animals↗

Comparison of semen quality obtained by vibratory stimulation and masturbation.

Six normal males underwent both penile vibratory stimulation and masturbation in order to compare the quantity and quality of semen produced by each method. There was no significant difference in the quantity and quality of the ejaculates produced by vibratory stimulation of the penis and masturbation. In addition, biochemical analysis of the seminal fluid collected by both procedures revealed similar values between all specimens for nine organic constituents, seven inorganic constituents and seven metabolic enzymes. None of the subjects demonstrated retrograde flow of semen. These findings indicate that vibratory stimulation is a 'physiological' means of inducing ejaculation, and can produce semen of normal quality.

Ejaculation↗

Osmo-sensitivity of the human sperm acrosome reaction.

The osmo-sensitivity of the human sperm acrosome reaction was investigated by determining the effect of hyper- and hypo-osmolal conditions on the ionophore A23187- and dbcAMP-induced reaction of both capacitated and non-capacitated spermatozoa. Hyper-osmolal conditions inhibited the agonist-induced reactions of both types of spermatozoa. Hypo-osmolal conditions caused a spontaneous loss of acrosomes from capacitated but not from non-capacitated spermatozoa. The loss of acrosomes under hypo-osmolal conditions was further enhanced by dbcAMP but not by ionophore A23187. Although significant decreases in sperm viability were occasionally observed at the high and low osmolalities, these decreases were not consistent and could not account for the observed loss of acrosomes. It is concluded that the human sperm acrosome reaction is osmo-sensitive. The acrosome reaction stimulated by ionophore A23187 (raises intracellular Ca2+) and dbcAMP (activates protein kinase A which causes protein phosphorylation) appears to involve water entry downstream from the action of these agonists. Preincubation in albumin (capacitation) causes human spermatozoa to lose their acrosomes under hypo-osmolal conditions. Finally, capacitation is not an essential prerequisite to the acrosome reaction as long as agonists are used that by-pass certain membrane-related events.

Acrosome↗

Absolute male factor infertility. A useful model for evaluating the efficacy of cryopreserved semen.

Cryopreserved semen generally results in diminished pregnancy rates when used for donor insemination. However, coexisting female subfecundity may significantly confound the pregnancy outcome. Therefore, in order to determine the efficacy of cryopreserved semen, 43 consecutive women with absolute male factor infertility (single women or wives of azoospermic men) were inseminated with cryopreserved spermatozoa using cervical caps. Life table analysis of the results revealed a cumulative probability of conception of 84% with a monthly fecundability rate of 19% after eight insemination cycles. These results are similar to those reported using fresh semen, suggesting that the efficacy of cryopreserved semen may be higher than previously believed.

Confounding Factors, Epidemiologic↗

Hemizona assay for measuring zona binding in the lowland gorilla.

The hemizona assay is a diagnostic test used to evaluate the binding potential of spermatozoa to zonae pellucida and has been used to predict fertilization potential in the human. In this study, frozen-thawed gorilla spermatozoa were coincubated with human hemizonae to evaluate tight binding and to assess the use of human zonae in evaluating sperm fertility. Matching hemizonae were incubated with human sperm to serve as a control. For evaluation of binding studies in a homologous system, matching halves of gorilla hemizonae were coincubated with both gorilla and human sperm. Whole, intact zonae of both human and gorilla oocytes were also coincubated with heterologous sperm to determine if penetration into the perivitelline space could occur. This study found that gorilla sperm bound well to both gorilla and human hemizonae, with a mean of 112.5 and 81.0 tightly bound sperm, respectively. Human sperm also bound to gorilla (mean 229.5) and human (mean 236.5) hemizonae. Following incubation with intact gorilla zonae, motile human sperm were found within the perivitelline space. However, gorilla sperm were not visible within the perivitelline space of nonviable human oocytes. These findings demonstrate that the zonae of nonviable human oocytes can be used to assess sperm binding of gorilla sperm. Studies continue for optimizing assay condition and correlation of findings with the fertility potential of gorilla sperm.

Animals↗

In vitro fertilization and gamete micromanipulation in the lowland gorilla.

PURPOSE: Our purpose was to utilize current reproductive technologies to treat an infertile female lowland gorilla. RESULTS: Following pituitary down-regulation with a gonadotropin releasing hormone agonist and follicular stimulation with human menopausal gonadotropins, transrectal ultrasound-guided aspiration of ovarian follicles yielded nine oocytes. Following failed fertilization with both epididymal and electroejaculated sperm, zona dissection and sperm injection were performed and produced one embryo, which was cryopreserved. Immature oocytes were obtained from ovarian sections and were either cultured in vitro or cryopreserved. CONCLUSIONS: This report demonstrates that in vitro fertilization techniques, including pituitary suppression, ultrasound-guided aspiration, micromanipulation, and cryopreservation, can be used to assist reproduction in the lowland gorilla.

Animals↗

Fertilizing capacity of various populations of spermatozoa within an ejaculate.

To determine whether ejaculates consist of various populations of sperm with varying degrees of fertilizing capacity, semen was filtered through a glass-wool column and the filtrate aliquoted sequentially into three portions. The ejaculate and its respective filtrates were evaluated for progressive motility, ability to respond to hypoosmotic swelling (HOS) test, acrosin content, ability to capacitate and acrosome react (AR), and ability to penetrate into denuded hamster oocytes (SPA). Filtrate 1 contained significantly more sperm with progressive motility, increased acrosin content, and positive HOS and SPA results than did the ejaculate. AR did not differ between the ejaculates and the respective filtrates. The data suggest that the ejaculate consists of various populations of sperm with different properties and that these populations may be separated by glass-wool filtration.

Acrosin↗